Nano glo cell reagent

Manufactured by Promega

Sourced in United States

The Nano-Glo cell reagent is a luminescent assay system designed for sensitive detection of enzymatic activities in live cells. It utilizes a proprietary luciferase enzyme and substrate to generate a luminescent signal proportional to the target enzyme's activity.

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Lab products found in correlation

Polyjet transfection reagent, by SignaGen (2 mentions) Synergy microplate reader, by Agilent Technologies (2 mentions) Passive lysis buffer (plb), by Promega (2 mentions) Synergy ht reader, by Agilent Technologies (2 mentions) X tremegene 9, by Roche (2 mentions) Quickchange lightning kit, by Agilent Technologies (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Furimazine, by Promega (1 mentions)

4 protocols using nano glo cell reagent

NanoLuc Luciferase Reporter Assay

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293T cells (3 × 10⁵ cells) were plated in 12-well plates in triplicate 24 h before transfection. Five hundred nanograms of the bioreporter constructs was transfected using PolyJet transfection reagent (SignaGen Laboratories). After 48 h, supernatant or cells lysates were collected. Cells were lysed using passive lysis buffer (Promega). NanoLuc luciferase assays were performed using one of two substrates: FMZ/ (Nano-Glo cell reagent, Promega) or native CTZ (3.33 μM final concentration; Nanolight Technologies-Prolume, Pinetop, AZ, USA). A Synergy microplate reader (BioTek, Winooski, VT, USA) was used to measure luminescence. Results are presented as RLU normalized to control. The data presented are the mean of three independent experiments.

Azad T., Singaravelu R., Taha Z., Jamieson T.R., Boulton S., Crupi M.J., Martin N.T., Brown E.E., Poutou J., Ghahremani M., Pelin A., Nouri K., Rezaei R., Marshall C.B., Enomoto M., Arulanandam R., Alluqmani N., Samson R., Gingras A.C., Cameron D.W., Greer P.A., Ilkow C.S., Diallo J.S, & Bell J.C. (2021). Nanoluciferase complementation-based bioreporter reveals the importance of N-linked glycosylation of SARS-CoV-2 S for viral entry. Molecular Therapy, 29(6), 1984-2000.

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MRAS-SHOC2 Protein-Protein Interaction Assay

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The NanoLuciferase reporter cell line (HEK293A-M2H-NLuc, tested negative for mycoplasma contamination) was generated by lentiviral transduction of a custom vector (pNGX_LV_017_GAL4_NLuc) bearing 5× GAL4 motifs upstream of a minimal promoter and the NanoLuciferase open reading frame (ORF). GAL4-DBD or VP16-TAD ORFs were cloned into custom lentiviral vectors (named pXP1510 and pXP1512) under an EF1a promoter and in frame with a ccdb cassette flanked by type IIS enzymes to enable golden-gate cloning of SHOC2 or MRAS cDNA (obtained by gene synthesis). Mutants were obtained by site-directed mutagenesis (Agilent QuickChange Lightning kit). HEK293A-M2H-Nluc cells were maintained in DMEM (Gibco) supplemented with 10% FBS (Gibco). Cells were plated at 1.25 × 10⁴ cells per well in a white transparent-bottom 96-well plates and transfected the next day with combinations of GAL4–MRAS and VP16–SHOC2 constructs using X-tremeGENE 9 (Roche) at a ratio 1:3 (DNA:transfection reagent). Then, 72 h after transfection, Nano-Glo cell reagent (Promega) was added and luminescence was measured using the Synergy HT reader (BioTek). The drawing in Fig. 3c illustrating the set-up of the M2H assay was generated using Biorender (www.biorender.com).

Hauseman Z.J., Fodor M., Dhembi A., Viscomi J., Egli D., Bleu M., Katz S., Park E., Jang D.M., Porter K.A., Meili F., Guo H., Kerr G., Mollé S., Velez-Vega C., Beyer K.S., Galli G.G., Maira S.M., Stams T., Clark K., Eck M.J., Tordella L., Thoma C.R, & King D.A. (2022). Structure of the MRAS–SHOC2–PP1C phosphatase complex. Nature, 609(7926), 416-423.

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NanoLuc Luciferase Assay for Biosensor Evaluation

Cited 1 time

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293T cells
(3 × 10⁵ cells) were plated in 12-well plates in triplicate
24 h before
transfection. An amount of 500 ng of the biosensor constructs was
transfected using PolyJet transfection reagent (SignaGen Laboratories).
After 48 h, supernatant or cells were collected. Cells were lysed
using passive lysis buffer (Promega), and NanoLuc luciferase assays
were performed using one of two substrates: furimazine, FMZ(Nano-Glo
Cell Reagent, Promega) or native coelenterazine, CTZ (3.33 uM final
concentration) (Nanolight Technologies – Prolume Ltd., Pinetop,
AZ, USA). Synergy Microplate Reader (BioTek, Winooski, VT, USA) was
used to measure luminescence. Results are presented as RLU (Relative
Luminescence Unit) normalized to control. The data presented are the
mean of three independent experiments.^{47 (link)}

Sadremomtaz A., Al-Dahmani Z.M., Ruiz-Moreno A.J., Monti A., Wang C., Azad T., Bell J.C., Doti N., Velasco-Velázquez M.A., de Jong D., de Jonge J., Smit J., Dömling A., van Goor H, & Groves M.R. (2021). Synthetic Peptides That Antagonize the Angiotensin-Converting Enzyme-2 (ACE-2) Interaction with SARS-CoV-2 Receptor Binding Spike Protein. Journal of Medicinal Chemistry, 65(4), 2836-2847.

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HEK293A Protein Interaction Assay

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HEK293A cells were plated at 1.25 × 10⁴ cells/well in a white transparent bottom 96-well plate and transfected the following day with LgBiT-PAX8 FL, PRDM3 FL-SmBiT, SmBiT-HNF1β, PCBD1-LgBiT, and PCBD1-SmBiT. X-tremeGENE 9 (Roche) was used for transfection with a ratio 1:3 (DNA:transfection reagent). At 48 h post transfection, Nano-Glo cell reagent (Promega) was added and luminescence was measured using a Synergy HT reader (BioTek).

Bleu M., Mermet-Meillon F., Apfel V., Barys L., Holzer L., Bachmann Salvy M., Lopes R., Amorim Monteiro Barbosa I., Delmas C., Hinniger A., Chau S., Kaufmann M., Haenni S., Berneiser K., Wahle M., Moravec I., Vissières A., Poetsch T., Ahrné E., Carte N., Voshol J., Bechter E., Hamon J., Meyerhofer M., Erdmann D., Fischer M., Stachyra T., Freuler F., Gutmann S., Fernández C., Schmelzle T., Naumann U., Roma G., Lawrenson K., Nieto-Oberhuber C., Cobos-Correa A., Ferretti S., Schübeler D, & Galli G.G. (2021). PAX8 and MECOM are interaction partners driving ovarian cancer. Nature Communications, 12, 2442.

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